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  本文關(guān)鍵詞：脾氣虛證患者唾液淀粉酶活性及其相關(guān)指標(biāo)的研究　出處：《廣州中醫(yī)藥大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 脾氣虛證 慢性淺表性胃炎 重癥肌無力 唾液 唾液淀粉酶活性

【摘要】：研究背景：既往研究根據(jù)“脾主涎”等中醫(yī)理論發(fā)現(xiàn)脾氣虛證患者唾液淀粉酶(salivary alpha-amylase, sAA)活性比值(酸刺激后／酸刺激前)較健康者明顯下降,該指標(biāo)1993年被衛(wèi)生部頒布的《中藥新藥臨床研究指導(dǎo)原則》列入脾氣虛證療效評價的參考指標(biāo),是為數(shù)不多的得到政府部門認(rèn)可的證候微觀指標(biāo)。但其在多種疾病脾氣虛證患者中的檢出率約為60%-70%,特異性不夠高,表明單一使用該指標(biāo)作為脾氣虛證的微觀指標(biāo)的信息還不夠全面。目的：以sAA活性改變?yōu)榍腥朦c,從同病異證和異病同證角度探討脾氣虛證患者sAA活性及其相關(guān)指標(biāo)(唾液pH值、唾液流率、總蛋白及離子濃度等)改變情況,進(jìn)一步豐富中醫(yī)“脾主涎”的科學(xué)內(nèi)涵,并觀察脾氣虛證患者sAA活性及相關(guān)指標(biāo)合參對臨床檢出率的影響。方法：一、運(yùn)用EP管自然流出法、口中轉(zhuǎn)動棉柱法和口中咀嚼棉柱法分別采集55例健康志愿者酸刺激前、后的唾液,檢測并比較sAA活性及總活性、唾液流率指標(biāo)變化情況,為篩選較優(yōu)的唾液采集方法提供依據(jù)。二、采集廣州中醫(yī)藥大學(xué)第一附屬醫(yī)院脾胃病科門診及住院部101例慢性淺表性胃炎(chronic superficial gastritis, CSG)和60例重癥肌無力(myasthenia gravis,MG)患者酸刺激前和酸刺激后的唾液,CSG患者納入的證型主要為脾氣虛、脾虛濕熱和肝胃不和3個證型(分別為36、40和25例),上午9：00～-11：00采集；MG患者納入的證型主要為脾氣虛、脾虛濕熱和脾腎兩虛3個證型(分別為32、23和5例),晨醒30 min內(nèi)采集。三、招募篩選青中年健康志愿者并采集其酸刺激前和酸刺激后的唾液,采集時間分晨醒30 min內(nèi)和上午9：00～11：00(早餐1h后)。四、檢測受試者sAA活性、唾液流率、pH值、總蛋白濃度、Ca2+及Cl-濃度。sAA活性采用Bernfeld法,唾液流率采用稱重法計算,pH值使用pH計檢測,總蛋白濃度采用BCA蛋白定量試劑盒檢測,Ca2+及C1-濃度采用離子定量試劑盒檢測。五、與健康對照組比較sAA活性及其相關(guān)指標(biāo)的改變情況,嘗試對sAA活性及相關(guān)指標(biāo)進(jìn)行多指標(biāo)合參并評價其臨床檢出率情況。結(jié)果：一、唾液樣品采集方法的優(yōu)化：EP管自然流取法與口中轉(zhuǎn)動棉柱法的sAA活性及總活性和唾液流率在酸刺激后均升高,反映了健康者唾液腺對酸的正常應(yīng)激能力,且兩者在酸刺激后各指標(biāo)升高的檢出率較為接近,其中又以口中轉(zhuǎn)動棉柱法有結(jié)果客觀、可平行采集足量唾液的優(yōu)勢；而口中咀嚼棉柱法的結(jié)果與前兩種方法差異較大,各指標(biāo)比值升高的檢出率明顯低于前兩者,未能客觀反映健康者酸刺激后sAA活性升高的現(xiàn)象。因而選擇口中轉(zhuǎn)動棉柱法作為后續(xù)研究唾液樣品的采集方法。二、脾氣虛證患者與健康者唾液指標(biāo)檢測：(一)CSG患者與健康者比較：(1)酸刺激后,健康者sAA活性、sAA總活性、流率和pH值升高,反映了健康者對酸刺激的正常應(yīng)激能力；唾液總蛋白濃度酸刺激后降低,可能是酸刺激后唾液流率升高,總蛋白濃度相對降低所致。(2)CSG及其脾氣虛證和脾虛濕熱患者酸刺激后sAA活性降低,并且sAA活性比值低于健康者,說明CSG及其脾虛患者唾液腺對酸的應(yīng)激能力降低,再次重復(fù)驗證既往研究結(jié)果。肝胃不和患者sAA活性及總活性比值高于脾氣虛證和脾虛濕熱證患者,與健康者無明顯差異；唾液流率、pH值、總蛋白濃度、Ca2+及C1-濃度在證型間無明顯差異；說明sAA活性改變?yōu)槠⑻摶颊呦鄬μ禺愋灾刚鳌?3)酸刺激前、后,CSG及其三證型患者唾液流率均低于健康者,總蛋白濃度、Ca2+及Cl-濃度均高于健康者,說明患者唾液成分濃度相對于健康者已發(fā)生改變,提示可能與患者植物神經(jīng)系統(tǒng)功能紊亂有關(guān)。(4)酸刺激后脾氣虛證患者唾液pH值降低的例數(shù)(16例)高于健康者(2例),說明脾氣虛證患者唾液緩沖能力降低。(二)MG患者與健康者比較：(1)MG及其脾氣虛證患者酸刺激后sAA活性降低,并且sAA活性比值和sAA總活性比值均低于健康者,說明MG及其脾虛患者唾液腺對酸的應(yīng)激能力降低,再次重復(fù)驗證既往研究結(jié)果。MG脾氣虛證患者sAA活性改變與CSG脾氣虛證患者表現(xiàn)類似,說明sAA活性改變?yōu)槠馓撟C患者普遍出現(xiàn)的現(xiàn)象。MG及其脾氣虛證和脾虛濕熱患者酸刺激前sAA活性高于健康者,提示在基礎(chǔ)狀態(tài)下MG脾虛患者交感神經(jīng)偏亢。 (2)MG及其脾氣虛證患者唾液流率酸刺激前后均低于健康者,總蛋白濃度、Ca2+及C1-濃度高于健康者,說明患者唾液成分濃度相對于健康者已發(fā)生改變,再次提示可能與患者植物神經(jīng)系統(tǒng)功能紊亂有關(guān)。(3)酸刺激后,脾氣虛證患者唾液pH值降低例數(shù)(17例)多于健康者(6例),pH值比值低于健康者,說明脾氣虛證患者唾液緩沖能力降低,這與CSG脾氣虛證患者表現(xiàn)類似,而MG脾氣虛證患者出現(xiàn)率更高,可能與MG病理改變相對更嚴(yán)重有關(guān)。(4)在脾氣虛證和脾虛濕熱證之間各指標(biāo)均無差異,提示可能是MG患者以脾氣虛弱為本,所以證型在指標(biāo)上未表現(xiàn)明顯差異。(5)CSG與MG脾氣虛證患者sAA活性比值均較健康者降低,重復(fù)證實了sAA活性比值降低可能是脾氣虛證患者的相對共性表現(xiàn),目前結(jié)果表明,兩種疾病脾氣虛證患者的唾液分泌可能存在相似的病理生理改變。結(jié)論：口中轉(zhuǎn)動棉柱法采集唾液樣品簡便、可靠,可供后期研究者繼續(xù)選用。本研究根據(jù)中醫(yī)理論“脾主涎”證實了CSG和MG脾氣虛證患者不僅僅表現(xiàn)在sAA活性異常這個單一的指標(biāo)上,還表現(xiàn)在sAA總活性、唾液流率、pH值、總蛋白濃度、Ca2+和Cl-濃度這些指標(biāo)改變上,為豐富“脾主涎”內(nèi)涵提供了參考。脾氣虛證和脾虛濕熱患者可能均存在植物神經(jīng)系統(tǒng)紊亂和唾液腺對酸的應(yīng)激能力降低的病理表現(xiàn)。除了sAA活性比值,唾液pH值比值改變對于脾氣虛證可能也有一定參考價值。將sAA活性、唾液流率和pH值進(jìn)行合參后在脾氣虛證的檢出率相較以往單用sAA活性指標(biāo)提高了約11.00%,但由于本研究的例數(shù)有限,多指標(biāo)合參的結(jié)果尚有待后續(xù)進(jìn)一步驗證。
[Abstract]:Background: Previous studies based on "spleen controlling saliva and saliva in patients with deficiency of TCM theory that temper amylase (salivary alpha-amylase, sAA) activity ratio (after acid stimulation / acid before stimulation) decreased significantly compared with the healthy people, the reference index in the 1993 by the Ministry of Health issued the" Chinese medicine clinical research guiding principles > included in evaluation of temper syndrome curative effect, is one of the few to get syndrome micro index recognition. But the government departments in a variety of diseases in patients with spleen deficiency syndrome, the detection rate is about 60%-70%, the specificity is not high enough, that single use of the index as microscopic indexes of spleen qi deficiency of the information is not comprehensive. Objective: to change the activity of sAA as the starting point, from the same angle with different syndromes syndrome and different disease activity and sAA in patients with deficiency of spleen index (pH value of saliva, salivary flow rate, total protein and ion concentration changes), To further enrich the scientific connotation of TCM "spleen controlling saliva", and to observe the activity of sAA in patients with deficiency of temper and related indicators and impact on the clinical detection rate. Methods: first, using EP tube natural outflow method, in turn chewing cotton fibre cotton column method and mouth were collected in 55 healthy volunteers acid after stimulation, saliva, detect and compare the activity of sAA and the total activity, changes of salivary flow rate index, provide the basis for screening the optimal saliva collection method. Two, collecting the First Affiliated Hospital of Guangzhou University of Chinese Medicine Department of outpatient and inpatient department and 101 cases of chronic superficial gastritis (chronic superficial, gastritis, CSG) and 60 cases of severe myasthenia gravis (myasthenia gravis, MG) patients before and after acid stimulation acid stimulated saliva syndrome CSG patients included mainly spleen deficiency, spleen deficiency and dampness heat and liver stomach disharmony syndrome (3 cases of 36,40 and 25 respectively), morning 9:00 - -1 1:00 collection; syndrome MG patients included mainly spleen deficiency, spleen deficiency and dampness heat and spleen kidney two empty syndrome type 3 (32,23 and 5), collected in the morning wake up 30 min. Three, the recruitment of young and middle-aged healthy volunteers were collected and screened the acid stimulation before and after acid stimulation of saliva collection time divided into morning wake up 30 min and 9:00 (1h to 11:00 morning after breakfast. Four), were determined by sAA activity, salivary flow rate, pH value, total protein concentration, Ca2+ concentration and Cl- activity of.SAA by Bernfeld method, the salivary flow rate by weight method calculation, pH value using the pH tester, total protein concentration. Detected by BCA protein assay kit, Ca2+ and C1- by ion concentration Quantification Kit detection. Five, change of control group sAA activity and the related indicators and health, try on the activity of sAA and related indicators of multi index parameters and to evaluate its clinical detection rate. Results: a saliva sample Optimization of acquisition method: EP tube natural flow method and rotation cotton column method in sAA activity and total activity and salivary flow rate were increased after acid stimulation, reflects the health of the normal salivary acid stress, and both increased in each index after acid stimulation rate is close to that of the and with the rotation of cotton column method is objective, parallel to capture enough saliva advantage; difference chew cotton column method with the former two methods greatly, each index ratio increased the detection rate was significantly lower than the former two, failed to reflect the healthy subjects after acid stimulation increased the activity of sAA and select the phenomenon. In turn the cotton column method as the collection method of follow-up study. Two saliva samples, detection of spleen qi deficiency patients and healthy subjects: (a) saliva index between CSG patients and healthy subjects: (1) after acid stimulation, healthy sAA activity, sAA activity, flow rate and P The increase of H value, reflects the health of the normal stress ability of acid stimulation; salivary total protein concentration decreased after acid stimulation, salivary flow rate may be increased after acid stimulation, total protein concentration is relatively reduced. (2) reduce the activity of sAA and CSG in spleen qi deficiency and spleen damp heat patients after acid stimulation, and sAA activity were lower than health, CSG and salivary glands of patients with spleen deficiency acid stress reduced ability to repeat past results. Verify the disharmony between liver and stomach in patients with sAA activity and total activity ratio was higher than that in patients with spleen deficiency syndrome and spleen deficiency and dampness heat syndrome, with no significant difference between healthy subjects; salivary flow rate, pH value, total protein concentration, Ca2+ had no significant difference. And the concentration of C1- in syndrome type; sAA activity change for patients with spleen deficiency relative specific indications. (3) after acid stimulation, CSG, and three patients were lower than that of normal salivary flow rate, total protein concentration, Ca2+ and Cl- The concentration was higher than that of healthy subjects, that saliva concentration patients compared to healthy have been changed, suggesting that it may be related to dysfunction of autonomic nervous system. (4) after acid stimulation of saliva in patients with deficiency of temper pH decreased the number of cases (16 cases) was higher than that of healthy subjects (2 cases), indicating a reduced capacity of saliva in patients with deficiency of buffer temper. (two) MG patients in comparison with healthy subjects: (1) reduce the activity of sAA and MG in patients with spleen deficiency syndrome after acid stimulation, and the activity of sAA and the ratio of the total activity of sAA was significantly lower than that of healthy subjects, MG and salivary gland in patients with spleen deficiency to acid stress reduced ability to repeat previous research results verify.MG temper the changes of activity of sAA and CSG in patients with syndrome of deficiency of spleen qi deficiency patients is similar to that of sAA activity is a common phenomenon in patients with spleen deficiency syndrome and spleen deficiency syndrome and.MG patients with spleen deficiency and dampness heat before acid stimulation of sAA activity is higher than that of healthy subjects Based on the state of MG, suggesting that spleen deficiency patients with sympathetic hyperactivity. (2) MG before and after acid stimulation in patients with spleen deficiency syndrome, salivary flow rate was lower than that of healthy subjects, the total protein concentration, Ca2+ and concentration of C1- was higher than that of healthy subjects, patients compared to healthy subjects saliva concentration has changed again, suggest that may be related with plant the disorder of the nervous system. (3) after acid stimulation, temper saliva in patients with deficiency of pH decreased the number of cases (17 cases) than healthy subjects (6 cases), pH ratio value is lower than that of healthy subjects, reduce the capacity of saliva in patients with deficiency of spleen and CSG buffer, the patients with spleen deficiency syndrome were similar, while MG appeared in patients with spleen deficiency syndrome at higher rates, and MG may be the pathological changes more serious. (4) between spleen deficiency syndrome and spleen deficiency syndrome of dampness heat of each index showed no difference, suggesting that MG patients with spleen weakness in the syndrome type, so in the index showed no significant difference Different. (5) sAA activity ratio of CSG and MG in patients with deficiency of spleen were lower than healthy subjects decreased, repeated that the lower sAA activity ratio may be relatively common symptoms of spleen deficiency syndrome patients, the results showed that two kinds of diseases in patients with spleen deficiency syndrome, the secretion of saliva may have a similar pathophysiological change. Conclusion: in rotation cotton column method to collect saliva samples is simple and reliable, can be used for later researchers to select the research. According to TCM theory of "spleen saliva" was confirmed by CSG and MG in patients with spleen deficiency syndrome, not only in the sAA abnormal activity of this single index, but also in the total activity of sAA, salivary flow rate, pH value, total protein concentration Ca2+ and Cl-, changing the concentration of these indicators, provide a reference for the rich connotation of "spleen controlling saliva". In patients with spleen deficiency syndrome and spleen deficiency and dampness heat may exist in autonomic nervous system disorders and salivary gland to acid stress ability decreased Pathology. In addition to the sAA activity ratio, the ratio of salivary pH change in spleen deficiency may also have certain reference value. The activity of sAA, salivary flow rate and pH of ginseng in spleen qi deficiency detection rate compared to the previous single sAA activity index increased by about 11%, but due to the limited number of research cases. Multi index combination results remains to be further verified.

【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R228

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